resDetect™ Human FGF2 ELISA Kit (Residue Testing)

背景（Background）

FGF basic (also known as FGF2 and HBGF-2) is an 18-34 kDa, heparin-binding member of the FGF superfamily of molecules (1-3). Superfamily members are characterized by the presence of a centrally placed beta -trefoil structure. FGF acidic (FGF-1) and FGF basic (FGF2) were the first two identified FGFs, and the designations acidic and basic refer to their relative isoelectric points. Human FGF basic is 288 amino acids (aa) in length. There are multiple start sites, four of which utilize atypical CUG codons, and one that initiates at an AUG start site (4 - 6). The four CUG start sites generate high molecular weight (HMW) FGF basic. There is a 34 kDa, 288 aa form, a 24 kDa, 210 aa form, a 22.5 kDa, 201 aa form, and a 22 kDa, 196 aa form. All are retained intracellularly, undergo extensive methylation, and possess one or more nuclear localization signals (NLS) (7-9). The AUG initiating form is 18 kDa and 155 aa in length. There is no signal sequence (ss). It is, however, secreted directly through the plasma membrane via a mechanism that appears to be dependent upon tertiary structure (10). In place of a ss, there is purportedly a 9 aa N-terminal prosegment that precedes a 146 aa mature segment (11). Early isolations of 18 kDa bovine FGF basic yielded 146 aa molecules, an effect attributed to the presence of acid proteases (12). The molecule contains a heparin-binding site (aa residues 128-144), and undergoes phosphorylation at Ser117 (13). There is also an ill-defined C-terminal NLS that may be more “functional” (or 3-dimensional) than structural (7). Human 146 aa FGF basic is 97% aa identical to mouse FGF basic (14).

应用说明（Application）

Human FGF2 ELISA Kit (Residue Testing) was developed for the detection and quantitative determination of GMP human FGF2 in samples from CAR-T product preparation processing.

It is for research use only.

重构方法（Reconstitution）

Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.

存储（Storage）

1. Unopened kit should be stored at 2℃-8℃ upon receiving.

2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.

3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

原理（Assay Principles）

This assay kit employs a standard sandwich-ELISA format, providing a rapid detection of Human FGF2. The kit consists of Pre-coated Anti-FGF2 Antibody Microplate and Human FGF2 Standard and Biotin-Anti-FGF2 Antibody and Streptavidin-HRP and buffers.

Your experiment will include 6 simple steps:

a) Bring all reagents to room temperature(20℃-25℃) before use.

b) Add your sample to the plate and take the Human FGF2 as standard. The samples and standard are diluted by Dilution Buffer.

c) Add the Biotin-Anti-FGF2 Antibody diluted by Dilution Buffer to the plate.

d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.

e) Wash the plate and add TMB.

f) Stop the substrate reaction by adding diluted acid. Absorbance (OD) is calculated by the absorbance at 450 nm minus the absorbance at 630 nm to remove background disturbance before statistical analysis. The OD Value reflects the amount of bound protein.